• 한국수산과학회지
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• 제39권1호
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• pp.23-26
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• 2006

Effects of sera from several fish species and insulin on the development of cultured Silurus asotus hepatocytes were investigated. Hepatocytes with high viability (95%) were obtained from the livers of male catfish by two step collagenase perfusion. Isolated hepatocytes, initially showed a typical round-shape, firmly attached to the culture dish within 24 h. In the presence of catfish serum, hepatocytes attached each other, spread well on the dish and developed into monolayer after 3-4 days of incubation. Cells within the established monolayer became polygonal in shape and their nuclei and boundaries being clearly visible under the microscope. In contrast, when incubated in FBS-supplemented or serum-free medium, cells managed to form small clusters, each made of 2-10 cells. Cells in FBS-supplemented medium further developed into larger clusters. However, these clusters failed to develope into monolayer. In addition, when insulin was deprived from culture medium, formation of monolayer also failed. From these data, it can be concluded that the presence of both catfish serum and insulin is necessary for the formation of monolayer of catfish hepatocytes and the functional role of fish serum may differ from that of insulin and can not be displaced by FBS-supplementation.

• Tissue Engineering and Regenerative Medicine
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• 제15권6호
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• pp.781-791
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• 2018

BACKGROUND: Glucosamine hydrochloride (GlcN HCl) has been shown to inhibit cell growth and matrix synthesis, but not with N-acetyl-glucosamine (GlcNAc) supplementation. This effect might be related to an inhibition of critical growth factors (GF), or to a different metabolization of the two glucosamine derivatives. The aim of the present study was to evaluate the synergy between GlcN HCl, GlcNAc, and GF on proliferation and cartilage matrix synthesis. METHOD: Bovine chondrocytes were cultivated in monolayers for 48 h and in three-dimensional (3D) chitosan scaffolds for 30 days in perfusion bioreactors. Serum-free (SF) medium was supplemented with either growth factors (GF) $TGF-{\beta}$ ($5ng\;mL^{-1}$) and IGF-I ($10ng\;mL^{-1}$), GlcN HCl or GlcNAc at 1mM each or both. Six groups were compared according to medium supplementation: (a) SF control; (b) SF + GlcN HCl; (c) SF + GlcNAc; (d) SF + GF; (e) SF + GF + GlcN HCl; and (f) SF + GF + GlcNAc. Cell proliferation, proteoglycan, collagen I (COL1), and collagen II (COL2) synthesis were evaluated. RESULTS: The two glucosamines showed opposite effects in monolayer culture: GlcN HCl significantly reduced proliferation and GlcNAc significantly augmented cellular metabolism. In the 30 days 3D culture, the GlcN HCl added to GF stimulated cell proliferation more than when compared to GF only, but the proteoglycan synthesis was smaller than GF. However, GlcNAc added to GF improved the cell proliferation and proteoglycan synthesis more than when compared to GF and GF/GlcN HCl. The synthesis of COL1 and COL2 was observed in all groups containing GF. CONCLUSION: GlcN HCl and GlcNAc increased cell growth and stimulated COL2 synthesis in long-time 3D culture. However, only GlcNAc added to GF improved proteoglycan synthesis.

• Biotechnology and Bioprocess Engineering:BBE
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• 제7권3호
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• pp.163-170
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• 2002

This report describes the use of a transtubular bioreactor to study the relative effects of diffusion versus perfusion of medium on antibody production by a hybridoma cell line. The study was performed with a high-density cell culture maintained in a serum-free, low-protein medium for 77 days. It was determined that the reactor possessed a macro-mixing pattern residence time distribution similar to a continuous stirred tank reactor (CSTR), However, due to the arrangement of the medium lines in the reactor, the flow patterns for nutrient distribution consist of largely independent medium path lengths ranging from short to long. When operated with cyclic, reversing, transtubular medium flow, some regions of the reactor (with short residence times) are more accessible to medium than others (with long residence times). From this standpoint, the reactor can be divided into three regions: a captive volume, which consists of medium primarily delivered via diffusion; a lapped volume, which provides nutrients through unilateral convection; and a swept volume, which operates through bilateral convection. The relative sizes of these three volumes were modified experimentally by changing the period over which the direction of medium flow was reversed from 15 min (larger captive volume) to 9 h (larger swept volume). The results suggest that antibody concentration increases as the size of the diffusion-limited (captive) volume is increased to a maximum at around 30 min with a sharp decrease thereafter. As reflected by changes in measured consumption of glucose and production of lactate, no significant difference in cellular metabolism occurred as the reactor was moved between these different states. These results indicate that the mode of operation of the transtubular bioreactor may influence antibody productivity under serum-free, low-protein conditions with minimal effects on cellular metabolism.

• Biotechnology and Bioprocess Engineering:BBE
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• 제1권1호
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• pp.32-35
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• 1996

Amoving aeration-membrane (MAM) bioreactor was employed for the production of 2$\mu$g/mL of tissue type Plasminogen Activator (tPA)in serum free medium from normal human fibroblast cells. This system could maintain high cell density for long periods of steady state conditions in perfusion cultivation. Under normal operating condition, shear stress was as low as 0.65 dynes/$\textrm{cm}^2$ at the agitation speed of 80 rpm. Even though cell density gradually decreased with increasing agitation speed, tPA production increased linearly with increasing shear stress within a moderate range. This culture system allowed production of 2$\mu$g tPA/mL while maintaining a high cell denisty of 1.0$\times$107 viable cell/mL.

• Journal of Chest Surgery
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• 제32권12호
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• pp.1119-1122
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• 1999

Native valve endocarditis (NVE) without preexisting structural valve or congenital cardiac malformation especially in pediatric group is rare. A case of isolated tricuspid valve endocarditis in a 7-year-old child without any cardiac malformation is described. This child had suffered from fever and productive cough for 3 weeks. Blood culture grew Staphylococcus aureus. Fever was not controlled even with proper antibiotic treatment. Transthoracic echocardiogram and lung perfusion scan revealed a large vegetation on the tricuspid valve with multiple embolism Surgical procedures included vegetectomy partial cusps resection and pericardial patch valvuloplasty. Th patient was in NYHA class I during follow up.

• Journal of Ginseng Research
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• 제18권1호
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• pp.17-24
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• 1994

Effect of ginsenoside mixture, ginsenoside $Rb_1$,$Rb_2$,$Rg_1$ and the fat soluble fraction of the roots of Panax ginseng C.A. Meyer on the activity of glucokinase (GK) in vitro has been observed and found that GK activity was increased about 15c1c at the concentration of ginsenoside mixture and/or the fat soluble fraction being $10^{-7}$,$10^{-5}$%. It was also observed that glucose uptake by rat liver was increased in the presence of either ginsenoside mixture or the fat soluble fraction by perfusion technique. Ginsenoside mixture stimulated various enzymes related to glucose metabolism, however, both ginsenoside mixture and the fat soluble fraction did not stimulate GK activity as expected. Primary culture of liver cells showed that the ginsenoside mixture and the fat soluble fraction increased GK activity significantly and they stimulated the GK activity synergistically in the co-presence of insulin.

• 한국미생물·생명공학회지
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• 제22권4호
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• pp.389-394
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• 1994

8 X 10$^{6}$(viable cells/ml) of maximum cell density and 9000(IU/ml) of $\gamma$-IFN production were obtained at 55(ml/hr) of a perfusion rate by cultivating HSF cells using a moving membrane aeration bioreactor. This system proves to be an efficient culture process by maintaning 90% of viable cells during the whole cultivation periods. The metabolic molar quotient of glucose to lactate was 0.81 for overall ranges of glucose consumed while the evolution of ammonia was not linearly related to the consumption of glutamine. Low molar conversion ratio was observed in low consumptions of glutamine and high molar conversion ratio in high comsumptions. It also shows that the glutamolysis plays important role in the steady state conditions by evolving larger quantities of ammonia than lactate. At the above of 50 rpm, which is the optimum agitation speed for this bioreactor, the cell growth was severely affected while the IFN production was less decrea- sed, maintaing 1.5 X 10$^{-3}$(IU/cell/day) specific IFN production rate. The cumulatvie $\gamma$-IFN production was 7.2 X 10$^{8}$(IU) for 70 days of the cultivation, which yields 1 X 10$^{7}$ (IU/day) of IFN production rate. Therefore, a commercial production of $\gamma$-IFN by this culture process can be achievable by maintaining the above IFN productivity in a scaled-up culture system.

• BMB Reports
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• 제43권4호
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• pp.245-249
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• 2010

GDH has been known to be related with hyperinsulinism-hyperammonemia syndrome. We have screened new drugs with a view to developing effective drugs modulating GDH activity. In the present work, we investigated the effects of a new drug, KHG26377 on glutamate formation and GDH activity in cultured rat islets. When KHG26377 was added to the culture medium for 24 h prior to kinetic analysis, the $V_{max}$ of GDH was decreased by 59% whereas $K_m$ is not significantly changed. The concentration of glutamate decreased by 50% and perfusion of islets with KHG26377 reduced insulin release by up to 55%. Our results show that KHG26377 regulates insulin release by inhibiting GDH activity in primary cultured islets and support the previous studies for the connection between GDH activity and insulin release. Further studies are required to determine in vivo effects and pharmacokinetics of the drug.

• Tuberculosis and Respiratory Diseases
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• 제44권5호
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• pp.975-991
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• 1997

배 경 : INH와 RFP에 동시내성을 보이는 다제내성 폐결핵은 적극적인 내과적 치료에도 불구하고 치료실패율이 40%에 이른다. 과거 폐결핵의 치료에 사용되었던 폐절제술이 다제내성 폐결핵의 보조적 치료수단으로 최근 다시 등장하였다. 방 법 : 1991년 1월부터 1995년 12월까지 HIV 감염이 없으면서 INH와 RFP에 동시내성을 보이는 다제내성 폐결핵 환자중 폐결핵 자체에 대한 보조적 치료로써 시행한 환자 14명을 대상으로 하여 후향적으로 의무기록과 방사선촬영소견 등을 조사하였다. 결 과 : 내과적 치료에도 불구하고 12명 (86%)의 환자에서 수술전 객담 균음전에 실패하였고 수술전 균음전이 이루어진 2명의 환자에서도 절제된 폐조직에서 실시한 결핵균 배양검사는 양성이었다. 흉부 전산화단층촬영상 양측성 병변이 10명(71%)에서 관찰되었다. 늑막폐절제술을 포함한 전폐절제술을 6명, 엽절제술 6명, 2명에서 시행하였다. 수술전 1초간 노력성 폐활량이 2L 이하였던 7명의 환자에서 수술전 폐관류스캔을 실시하였고, 절제된 폐로의 관류 중앙값 4.8%에 불과하였다. 수술과 관련한 사망은 없었으며, 1명에서 수술후 폐늑막루를 포함한 농흉이 발생하였고, 1명에서 3주이상 흉관을 통한 공기유출이 있었다. 수술후 13명(93%)의 환자에서 균음전화가 이루어졌고 중앙값 23개월간의 추적관찰기간동안 재발은 없었다. 1명(7%)의 환자는 폐절제술후에도 객담 균양성이 지속되었다. 결 론 : 과거 국내외에서 보고된 다제내성 폐결핵의 내과적 치료성적과 비교해 볼 때, 병변이 국한되어있고 적절한 폐기능을 가진 다제내성 폐결핵 환자에서 실시되는 보조적인 폐절제술은 중요한 역할을 할 것으로 기대된다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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• pp.425-428
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• 2000

본 연구에서는 재조합단백질 생산을 위한 식물세포배양용 세포배양기 운전에서 배지교환식 배양과 기체조성에 따른 영향을 관찰하였다. 배지교환식 배양에서는 6일째에 배지교환을 시작하는 것이 세포의 생장을 저해하지 않았으며 교환용 배지의 조성을 120 g/L sucrose와 2배 농축된 염농도를 하였을 때 가장 높은 세포농도인 20.0 gDCW/L를 나타내었으며 GUS 생산량은 60 g/L sucrose와 기본배지와 같은 농도의 염농도로 배지를 교환했을 때 12.9 U/mL로 다른 경우에 비하여 $1.5^{\sim}4.6$ 배 증가된 결과를 보였다. 그리고 $CO_2$의 농도를 5%로 높인 공기로 통기했을 때는 세포의 초기생장을 증진시켰으며 0.2 vvm의 통기량에서 5% $CO_2$로 통기시킨 경우는 일반 공기를 0.1 vvm으로 통기시킨 경우보다도 77% 높은 GUS의 생산량을 나타내었다.